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Lariat capping as a tool to manipulate the 5' end of individual yeast mRNA species in vivo.

RNA. 2017; 
Krogh N, Pietschmann M, Schmid M, Jensen TH, Nielsen H.
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摘要

The 5' cap structure of eukaryotic mRNA is critical for its processing, transport, translation, and stability. The many functions of the cap and the fact that most, if not all, mRNA carries the same type of cap makes it difficult to analyze cap function in vivo at individual steps of gene expression. We have used the lariat capping ribozyme (LCrz) from the myxomycete Didymium to replace the mRNA m7G cap of a single reporter mRNA species with a tiny lariat in which the first and the third nucleotide are joined by a 2', 5' phosphodiester bond. We show that the ribozyme functions in vivo in the budding yeast Saccharomyces cerevisiae presumably without cofactors and that lariat capping occurs cotranscriptionally. T... More

关键词

lariat capping ribozyme; m7G; mRNA processing; mRNA turnover; polyadenylation